摘要
由于近年来不可治愈疾病的增加和COVID-19的流行，反义2′-fluoro修饰寡核苷酸药物和mRNA疫苗封顶引物引起了全球关注。为了确保稳定和经济地生产这些寡核苷酸和引物，必须建立可靠的方法，以从相关反应的产物中获得大量高纯度(＞99%)的相应单体(2′-fluoro核苷亚磷酰胺，下文缩写为“2′F-NP”)。目前，该任务已经使用基于分批(非连续)模式的两阶段分离过程(串联萃取和色谱)来处理，然而，这种方法存在溶剂消耗过多和产率低的问题。2′F-NP的清洁生产需要解决这些问题。为了解决这一问题，本研究旨在开发一种新的一步法，即可连续回收高纯度的2′F-NP。为此，我们将模拟移动床(SMB)方法引入到目标2′FNP回收工艺的开发阶段，并进一步建立了合适的SMB结构和溶剂梯度策略，以连续同时去除所有杂质。然后使用估计的固有参数、探索的再生方法和准备的优化工具优化基于这些考虑确定的SMB模式过程。进行了相关的SMB实验，证实了本研究中开发的工艺在不借助进一步工艺的情况下以高纯度(99.5%)和高收率(96.5%)连续模式回收2′F-NP方面是相当有效的，从而能够显著降低溶剂消耗。本文的研究结果将有助于2′F-NP的清洁、环保生产，也将有助于反义寡核苷酸药物和rna疫苗封顶引物的生产。
Abstract
Due to the recent increase in incurable diseases and the pandemic of COVID-19, there have been global interests in antisense 2′-fluoro modified oligonucleotide drugs and mRNA-vaccine capping primers. To ensure stable and economical production of these oligonucleotides and primers, it is essential to establish a reliable process for securing a large amount of their corresponding monomer (2′-fluoro nucleoside phosphoramidite, abbreviated as “2′F-NP” hereafter) at high purity (＞99%) from the output of the relevant reaction. Currently, this task has been handled using two-stage separation processes (extraction and chromatography in series) based on batch (noncontinuous) modes, which, however, had the problems of excessive solvent consumption and low yield. For cleaner production of 2′F-NP, these problems should be resolved. To address such issue, this study aimed to develop a new single-step process that can recover 2′F-NP at once in a continuous mode at high purity. For this purpose, we introduced a simulated-moving-bed (SMB) method into the development phase of the targeted 2′FNP recovery process, and further established the appropriate SMB configuration and solvent-gradient strategy to continuously remove all the impurities at once. The SMB mode process determined based on such considerations was then optimized using the intrinsic parameters estimated, the regeneration method explored, and the optimization tool prepared. The relevant SMB experiment was carried out, which confirmed that the developed process in this study was quite effective in continuous-mode recovery of 2′F-NP at high purity (99.5%) and high yield (96.5%) without the aid of a further process, thereby enabling a significant reduction in solvent consumption. The results in this paper will thus contribute to the clean and eco-friendly production of 2′F-NP, which will also be of benefit to antisense oligonucleotide drug and mRNA-vaccine capping primer production processes.
NETL
NSTL
Elsevier